Derivatives of zearalane

ABSTRACT

wherein A is a member selected from the group consisting of hydrogen, hydroxy, and -OR; R is a member of the group consisting of hydrogen, lower alkyl, aralkyl, lower alkanoyl and benzol; B is a member of the group consisting of hydrogen and nitro, provided that B cannot be nitro when A is hydrogen; and R&#39;&#39; is a member of the group consisting of -CH2OH, -CHO, COOR&#39;&#39;&#39;&#39; and CH2COOR increment wherein R&#39;&#39;&#39;&#39; is a member of the group consisting of hydrogen, lower alkyl and aralkyl, are administered orally and parenterally to animals in order to achieve an anabolic and estrogenic response such as increasing the growth rate of meat-producing animals.   D R A W I N G

United States Patent [72] Inventors Norman P. Jensen Watchung; Thomas B.Windholz, Westfleld, both of NJ. [21 Appl. No. 803,048 [22] Filed Feb.27, 1969 [45} Patented Nov. 16, 1971 [73] Assignee Merck & Co., Inc.

Rahwly, NJ.

[ 54] DERIVATIVES 0F ZEARALANE 17 Claims, No Drawings 52] us. Cl...26o/343.2'F, 424/279 [5 l 1 Int. Cl C07d 9/00 [50] Field oiSearch260/343.2F

[56] References Cited UNITED STATES PATENTS 3,373,026 3/ l968 Hodge etal 260/3431 3,373,039 3/1968 Hodge et a1 260/3432 X PrimaryExaminer-John M. Ford Attorneys-l. Louis Wolk, J. Jerome Behan and F. M.Mahon nasrmcr:

wherein A is a member selected from the group consisting ofhydrogemhydroxy, and OR; R is a member of the group consisting ofhydrogen, lower alkyl, aralkyl, lower alkanoyl and benzol; B is a memberof the group consisting of hydrogen and nitro, provided that B cannot benitro when A is hydrogen; and R is a member of the group consisting ofCH,OH, CHO, COOR" and CH,COOR wherein R" is a member of the groupconsisting of hydrogen, lower alkyl and aralkyl, are administered orallyand parenterally to animals in order to achieve an anabolic andestrogenic response such as increasing the growth rate of meat-producinganimals.

DERIVATIVES F ZEARALANE This invention relates to new and usefulcompositions of matter classifiable in the field of organic chemistry as7'- hydroxymethyl, 7-formyl, 7'-carboxy and 7'-carbomethoxymethylderivatives of uaralane 6-( IO-hydroxyundecyD-B- resorcyclicacid-u-lactone. More particularly, this invention relates to7'-hydroxymethyl, 7'-formyl, 7'-carboxy and 7'-carbomethoxymethylderivatives of zearalane and to 2-mono-, 4- monoand 2,4-dietherderivatives thereof; to 2-mono-, 4- monoand 2,4-diester derivativesthereof; to 2-desoxy and 5- nitro derivatives thereof; to methods ofpreparing such compounds and to the use of such compounds as estrogenicand anabolic agents.

The novel compounds of this invention may be represented by thefollowing structural formula:

wherein A is member selected from the group consisting of hydrogen,hydroxy and OR; R is a member of the group consisting of hydrogen,loweralkyl, aralkyl, loweralkanoyl and benzoyl; B is a member of thegroup consisting of hydrogen and nitro, provided that B cannot be nitrowhen A is hydrogen; and R is a member of the group consisting of -CH,OH,-CHO, -COOR" and -CH -COOR" wherein R" is a member selected from thegroup consisting of hydrogen, loweralkyl and aralkyl. it will beappreciated by those skilled in the art that the carbon atom at the7'-position in the above fonnula is asymmetric and that there are,therefore, two diasterioisomers of the compounds of this invention.

The instant invention is based upon applicants discovery that thecompounds described above display valuable anabolic and estrogenicactivity. It is contemplated that dosage units containing thesecompounds as the essential active ingredient will be administered eitherorally or parenterally to animals to achieve an anabolic and esterogenicresponse such as increasing the rate of growth of meat producinganimals.

As used herein, the terms, loweralkyl and loweralkanoyl, are intended toinclude both straight and branched-chain loweralkyl and loweralkanoylgroups having from one to six carbon atoms in the alkyl moiety. Typicalof such groups are, for example, methyl, ethyl, propyl, isopropyl,butyl, n-hexyl, formyl, acetyl, isopropionyl, n-butyryl, and the like.The preferred aralkyl group is benzyl.

As the starting material in the preparation of the novel zearalanes ofthis invention, applicants employ zearalenone 2,4-dibenzyl ether. Thisis a well-known compound and is readily obtained from zearalenone byethen'fication techniques already fully described in the art.

Applicants have found that zearalenone 2,4-dibenzyl ether readily can beconverted into 7'-fonnylzearalane via a fourstep synthesis which firstinvolves treatment of zearalenone 2,4dibenzyl ether with ethyl fonnatein the presence of a strong base such as, for example, sodium hydride,alkali and alkaline-earth metal hydroxides, sodamide, alkali metalalkoxides, and the like in order to produce7-hydroxymethylenezearalenone 2,4-dibenzyl ether (which compound, ofcourse, may exist also in tautomeric fonn as 7-formylzearalenone2,4-dibenzyl ether. The reaction proceeds satisfactorily at about roomtemperature and is carried out in a suitable dry organic solvent suchas, for example, benzene, toluene, ethylene glycol dimethyl ether,hexane and the like. Usually, from about 12 to about 24 hours isrequired for completion. Applicants have found that the reaction is mostsatisfactorily carried out using sodium hydride as the base and benzeneas the solvent. After decomposition of any excess sodium hydride in thereaction mixture by the addition of water, the product may be recoveredfrom an ether extract in the conventional manner. Preferably, theproduct is employed in the next stage of the synthesis without furtherpurification.

7'-Hydroxymethylenezearalenone 2,4-dibenzyl ether as obtained above isthen treated with an aliphatic alcohol, preferably with cyclohexanol, toform the corresponding 7'- ether. Thus, 7'-hydroxymethylenezearalenone2,4- dibenzylether may be treated with cyclohexanol to prepare 7'-cyclohexyloxymethylenezearalenone 2,4-dibenzyl ether. The reaction iscarried out in a dry organic solvent, benzene being preferred, at refluxtemperature and usually requires from about l2 to about 48 hours forcompletion. Desirably, the reaction is carried out in the presence of acatalytic quantity of a strong organic acid such as, for example,p-toluene sulfonic acid and trifluoroacetic acid. The product obtainedby removal of volatiles from the reaction mixture may be purified byconventional column chromatography using, for example, silica gel andeluting with solvent mixtures such as 10 percent ethyl acetate inbenzene.

The 7'-ether intermediate obtained above is then converted into6'(7')-dehydro-7'-formylzearalene 2,4-dibenzyl ether by treating the7'-ether with a reducing agent such as, for example sodium borohydrideor lithium-aluminum hydride followed by treatrnent of residue from thisreaction with a strong mineral acid such as hydrochloric and sulfuricacids. Both reactions are carried out in a suitable organic solvent suchas methanol, ethanol ether, benzene and the like and both may be carriedout at room temperature. The 6(7)-dehydro-7'- formylzearalene2,4-dibenzyl ether so obtained may be purified by conventional columnchromatography as indicated above.

7'-Formylzearalane then is prepared by subjecting 6'(7')-dehydro-7'-formylzearalene 2,4-dibenzyl ether to conventional catalytichydrogenation. Applicants have found that this reduction selectivelysaturates the double bonds at the l'(2')- and 6'(7')-positions withoutsaturation of the benzenoid moiety and simultaneously cleaves thedibenzyl ether groups at positions 2 and 4. The reaction may be carriedout at atmospheric pressure and at ambient temperatures in a suitableorganic solvent, such as benzene and ethyl acetate, and in the presenceof a conventional hydrogenation catalyst such as 10 percentpalladium-on-carbon. The desired 7'-formylzearalane is recovered fromthe reaction mixture, after separation of the catalyst, by removal ofthe solvent and is purified by column chromatography over silica gelusing 10-20 percent ethylacetate in benzene as the eluant.

As indicated above, the carbon torn at the 7'-position in 7'-formylzearalane is asymmetric. This compound, therefore, exists in twodiastereo isomeric forms. it will be understood that the7'-forrnylzearaland as obtained in the reactions described above, isobtained as a mixture of the two isomeric forms. Applicants have foundthat the isomers may be separated by fractional crystallization frombenzene. The least soluble isomer, isomer A, separates upon cooling toroom temperature from a hot saturated solution of the epimeric mixturein benzene. The more soluble isomer, isomer B, is precipitated from thebenzene filtrate obtained after separation of isomer A by dilution' ofthe filtrate with hexane. Both isomers may be purified by chromatographyover silica gel followed using l020per cent ethyl acetate in benzene aseluant. Recrystallization from benzene yields pure isomer A (m.p.l40-l42 C.) and pure isomer B (m.p. l48-l52 C.).

Preparation of those 7-formylzeara.lanes of this invention which have analkoxy or aralkoxy group at the 2- and 4-positions may be achievedreadily according to techniques already fully described in the art bytreating 7'-formylzearalane with a dialkylor diaralkyl sulfate in anaqueous basic medium. Thus, for example, 7'-formylzearalane in aqueoussodium hydroxide may be treated with dimethyl sulfate to prepare2,4-dimethoxy-7'-formylzearalane. The reaction may be carried out attemperatures ranging from about 15 C. to about room temperature and isusually complete in about 30 minutes to 2 hours. The precipitateddiether is recovered by filtration and is purified by recrystallizationin the conventional manner.

It has been found also that upon acidification of the filtered reactionmixture obtained above, there is obtained a second precipitated productwhich may be recovered by filtration and purified in the usual way. Thissecond product is the 2- monoalkyl (or Z-monoaralkyl) ether of7'-formylzearalane. The reaction described above, therefore, affords aroute to both the 2,4-diethers and 2-monoethers of zearalane, it will beunderstood that in order to prepare any particular 2,4-diether or2-monoether it is necessary merely to employ the appropriate dialkyl ordiaralkyl sulfate as the alkylating reagent.

The 7-formylzearalanes of this invention having a 4- monoalkyl ether ora 4-monoaralkyl ether group conveniently are prepared from thecorresponding 2,4-diether by selective hydrolysis. This may be effectedby treating the diether in a suitable organic solvent such as methylenechloride with either boron tribromide or boron trichloride at about 10to about -l5 C., preferably under an inert atmosphere such as nitrogen.The hydrolysis of the ether radical at the 2-position is essentiallyinstantaneous and, accordingly, the diether is contacted onlymomentarily with the boron trihalide, i.e., preferably for less than lminute. The 4-monoether is isolated by removal of the solvent andrecrystallization from a suitable organic solvent such as nitromethane.

7'-formylzearalanes of this invention having a loweralkanoyloxy orbenzoyloxy substituent at the 2- and 4-positions may be prepared bysubjecting 7'-formylzearalane to the action of an acylating agent.Conveniently, the acylation is carried out by treating the7'-formylzearalane, dissolved in a suitable organic solvent such aspyridine, with an acid anhydride. Thus a pyridine solution of7'-formylzearalane may be treated with acetic anhydride in order toobtain 2,4- diacetoxy-7-formylzearalane. The reaction may be carried outat room temperature and usually is complete in 4-12 hours. The diestermay be recovered from the reaction mixture by pouring the reactionmixture over ice, extracting the resulting mixture with ether, dryingthe extract and conventional recrystallization. It will be understoodthat it is necessary merely to select the appropriate acid anhydride asthe acylating agent in order to prepare any of the diesters of thisinvention.

Those of the 7-formyzearlanes of this invention having amonoloweralkanoyl or a monobenzoyl ester grouping at the 2- or4-positions may be prepared by subjecting a 2-monoether or 4-monoetherderivative of 7-formylzearalane to acylation according to the techniquedescribed above in order to prepare the corresponding2-monoether-4-ester or 2-ester- 4monoether derivative. These mixedether-ester derivatives which also are included within the scope of thisinvention then may be subjected to boron trihalide hydrolysis asdescribed above in order to obtain the desired 2- or 4-monoesterderivative.

Mixed 2,4-diether and 2,4-diester derivatives of 7-formylzearalane arereadily prepared from a 2- or 4-monoether or from a 2- or 4-monoester bythe etlierification or esterification techniques described above. Thusfor example, 2-methoxy-4- ethoxy-7'-formylzearalane may be prepared bytreating 2- methoxy-7-zearalane with diethylsulfate and 2-acetoxy-4-propionyloxy-7'-formylzearalane may be prepared by treating2-acetoxy-7'-formyl-formylzearalane with propionic anhydride accordingto the techniques already fully described.

It will be understood that where it is desired to obtain any of theether or ester derivatives described above in their individual isomericforms, it is necessary only to subject an epimeric mixture to fractionalcrystallization techniques, such as described above, or to employ thedesired 7'-formylzearalane isomer as the starting material in theetherification or esterfication reaction. The 7'-formylzearalanesdescribed above, together with their ether and ester derivatives,display valuable anabolic and estrogenic activity and serve also as keyintermediates in the preparation of the other novel zearalanes of thisinvention.

The 7'-carboxyzearala.nes of this invention may be prepared by oxidationof 7'-formylzearalane with standard Jones Reagent 26.7 g. of chromiumtrioxide in 23 ml. of concentrated sulfuric acid brought to a volume ofml. with water). The reaction is carried out in a suitable organicsolvent such as, for example acetone, benzene, chloroform and the like,at temperatures ranging between -20 C. to 10 C. Even at these lowtemperatures, the reaction proceeds rapidly and usually is complete inform about 5 to about 20 minutes. The 7'-carboxyzearalane so producedmay be purified by conventional thinlayers chromatography on silica gelplates. Ether and ester derivatives of 7'-carboxyzearalane may beprepared by the esterification and etherification processes describedabove or by selecting the appropriately substituted 7'-formylzearalaneas starting material.

The 7'-hydroxymethylzearalanes of this invention may be obtained bytreating 7'-carboxyzearalane with a reducing agent such as, for example,sodium borohydride and lithiumaluminum hydride. The reaction proceedssatisfactorily at room temperature in an organic solvent such as, forexample, methanol, ethanol, dioxane and the like and is usually completein from about i to about 2 hours. Removal of the solvent in vacuofollowed by ether extraction yields a crude residue which may bepurified by conventional thin-layer chromatography. Ether and esterderivatives of 7'-hydroxymethylzearalane may be prepared by thetechniques described above or by employing an appropriately substituted7-carboxyzearalane as the starting material.

Applicants have found that the side chain in the 7-position of the7'-carboxyzearalanes of this invention may be extended by one carbonatom to obtain the corresponding 7-carboxymethylzearalanes and thatthese compounds, together with their 7'-carbomethoxyloweralkyl (andaralkyl) ester derivatives, display the same anabolic and estrogenicactivity as the 7'-carboxyzea.ralanes. These 7-carboxymethylzearalanesmay be prepared by treating 7-hydroxymethylzearalane or an ether orester derivative thereof, such as described above withp-toluenesulfonylchloride in a dry organic solvent such as pyridine toform the corresponding 7'-p-toluenesulfonyloxymethylzearalane which,after isolation from the reaction mixture as a crude, is treated with analkali metal cyanide in a solvent such as dimethylformamide or ethyleneglycol dirnethyl ether to form the corresponding7'-cyanomethylzearalane. This compound also may be isolated from thereaction mixture as a crude and, without further purification, treatedwith a lower alkanol (or aralkanol) in the presence of gaseous hydrogenchloride to form the corresponding zearalane 7'- acetimidic acidloweralkyl (or aralkyl) ether which then may be treated in situ withwater to form the desired 7'-carboalkoxy( or aralkoxy)methylzearalane.The corresponding 7'-carboxymethylzearalanes are obtained readily byconventional basic hydrolysis of the preceding 7-ester.

The novel zearalanes of this invention having a nitro group in the5-position are prepared by treating the parent zearalane, or an ether orester derivative thereof, with concentrated nitric acid. The reactionpreferably is run at a temperature of about 10 C. and is usuallycomplete in from about 30 minutes to 2 hours. The crude Snitro zearalaneis separated from the reaction mixture by filtration and may be purifiedby conventional recrystallization.

Preparation of the Z-desoxy zearalanes of this invention may be achievedby treating 7-formylzearalane with commercial dihydropyran to form7'-formylzearalane 4- tetrahydropyranyl ether which then may be treatedwith lphenyl-S-chlorotetrazole to form the corresponding7'-formylzearalane 2-( l-phenyltetrazolyl) ether-4-tetrahydropyranylether. Applicants have found that catalytic hydrogenation of thisdiether followed by treatment with dilute mineral acid results incleavage of oxygen function at the 2-position with simultaneous cleavageof the ester group at the 4-position and reduction of the 7-formyl groupto hydroxymethyl yielding 2- desoxy-7'-hydroxymethylzearalane.

Those 2-desoxy zearalanes of this invention having a formyl group or acarboxy group in the 7-position are prepared by oxidation of7'-hydroxymethylzearalane with Jones Reagent according to the techniquesdescribed above. One-quarter milliliter of standard Jones Reagent isrequired for conversion of each millimole of 7,'-hydroxymethylzearalaneto 7'-formylzearalane and k milliliter of this reagent is required forconversion of each millimole of the hydroxy methyl compound to7-carboxyzearalane.

Preparation of 2-desoxy zearalanes having an ether or ester group in the4-position may be carried out by subjecting 2- desoxy-7'-formylzearalaneto the conventional etherification or esterification proceduresdescribed above. These 2-desoxy- 7'-formylzearalane ethers or esters maythen be converted to the corresponding 7'-hydroxymethyl compound byreduction (with sodium hydride, for example) or to the corresponding7'-carboxy compound by oxidation (with Jones Reagent) according to theprocedures previously described.

As pointed out above, the novel zearalanes of this invention displayvaluable anabolic and estrogenic activity and may be used to achieve ananabolic and esterogenic response such as increasing the rate of growthof meat producing animals. These compounds can be administered toanimals by any suitable method including oral and parenteraladministration. They may be blended with ordinary animal feedscontaining the usual nutritionally balanced diet of carbohydrates,proteins. vitamins and minerals in an amount sufficient to produce thedesired rate of growth and thus can be fed directly to the animals.Also, if desired, the compounds of this invention may be suspended in asuitable injectable suspension medium and injected parenterally. Theamount of compound administered to the animal, of course, will varydepending upon the animal and the desired rate of growth. Usually,administration of from about 5 to about 100 mg. per animal per dayresults in significantly increased growth rate.

The best mode contemplated by applicants for carrying out theirinvention is set forth in the following examples; it being understoodthat the examples are included for purposes of illustration merely.

EXAMPLE l 7'-Hydroxymethylenezearalenone 2-4,Dibenzyl Ether Treat 19 g.of zearalenone 2,4-dibenzyl ether with 4.2 g. of 54 percent sodiumhydride/mineral oil in 300 ml. of dry benzene and 1 ml. of dry t-butanolfollowed by 9.6 ml. of ethyl fonnate. Stir for 18 hrs. at roomtemperature and add an additional 3 ml. of ethyl forrnate. Continuestirring at room temperature for 3 hrs. and add 7 ml. of water todecompose the mixture. Isolate the product by acidifying the waterlayer, extracting the mixture with ether. Dry the ether extract andconcentrate to a gummy residue.

Dissolve 14.5 g. of the product obtained above in a mixture of 3.6 ml.of cyclohexanol, 50 mg. of p-toluene sulfonic acid and 350 ml. ofbenzene and reflux for 45 hrs. using a water separator filled withmolecular sieves for water removal. Cool the reaction mixture and washwith 1000 ml. of saturated brine containing 100 ml. of 2.5 N sodiumhydroxide. Dry the residue with magnesium sulfate and concentrate to g.of oil. Chromatograph the oil on silica gel using 10 percent ethylacetate in benzene to obtain pure 7-cyc1ohexyloxymethylenezearalenone2,4-dibenzyl ether.

EXAMPLE 2 6'( 7' )-Dehydro-7'-Formylzearalene 2,4-Dibenzyl Ether Add 2.5g. of sodium borohydride to a solution of 5.7 g. of7'-cyclohexyloxymethylenezearalenone 2,4-dibenzyl ether in 1 10 ml. ofmethanol. Stir for 1.5 hrs. and remove the solvent in vacuo. Take up theresidue in a mixture of 200 ml. of ether and 200 ml. of water. Separatethe ether layer and washed with saturated brine, dry with magnesiumsulfate and concentrated to 5.5 g. of residue. Stir the residue undernitrogen with ml. of ether and 100 ml. of 3 N. hydrochloric acid for 4hrs. Add 200 ml. of ether. Separate the ether layer, wash with saturatedbrine, dry with magnesium sulfate and concentrate to 5 g. of residue.Chromatograph the residue on silica gel using 10 percent ethyl acetatein benzene to obtain pure 6'(? ')-dehydro-7'-formylzearalene2,4-dibenzyl ether.

EXAMPLE 3 7'-Formylzearalane Hydrogenate 2.2 g. of6'(7')-dehydro-7'-formylzearalene 2,4-dibenzyl ether in 50 ml. of ethylacetate at atmospheric pressure over 10 percent palladium on carboncatalyst. When the theoretical amount of hydrogen is taken up, stop thereaction and remove the catalyst by filtration. Remove the solvent invacuo and chromatograph the residue over silica gel using 10-20 percentethyl acetate in benzene to obtain pure 7'-formylzearalane.

Dissolve the crude product obtained above, prior to chromatography, inhot benzene to form a saturated solution. Cool to room temperature andseparate the insolubles to obtain crude 7'-formylzearalane isomer A.Dilute the filtrate with hexane and separate the insolubles to obtain7'-formylzearalane isomer B. Purify both isomers by chromatography oversilica gel eluting with 10 percent ethyl acetate in benzene. Evaporatethe eluate to dryness in vacuo and recrystallize the residue frombenzene to obtain isomer A (m.p. l40-142 C.) and isomer B (m.p. 148-152C.).

EXAMPLE 4 7-Carboxyzearalane Dissolve 291 mg. of 7'-formylzearalane in15 m1. of acetone and cool to 0 C. Add 0.25 ml. of standard JonesReagent (26.7 g. of chromium trioxide in 2.3 m1. of concentratedsulfuric acid brought to 100 ml. with water) and mix for 8 min. Add 5ml. of methanol to decompose the mixture and remove the solvent invacuo. Take up the residue in a mixture of 20 ml. of water. Separate theether layer, wash with saturated brine, dry with magnesium sulfate andconcentrate to 0.3 g. of residue. Purify by chromatography on silica gelplates using 10 percent ethyl acetate in benzene to obtain7'-carboxyzearalane.

EXAMPLE 5 7-l-lydroxyrnethylzearalane Dissolve 340 mg. of7'-formy1zearalane in 10 ml. of methanol and add 0.26 g. of sodiumborohydride in small portions with stirring over a period of 2 mins.Stir at room temperature for 1.25 hours and remove the solvent in vacuo.Take up the residue in a mixture of 30 ml. of ether and 30 ml. of 5percent hydrochloric acid. Separate the ether layer, wash with saturatedbrine, dry with magnesium sulfate and concentrate to dryness to obtain7'-hydroxymethy1zearalane. Purify by thin -layer chromatography onsilica gel plates using 10 percent ethyl acetate in benzene as theeluant to obtain 7'- hydroxymethylzearalane.

EXAMPLE 6 7-Carbomethoxymethylzearalane Dissolve 0.34 g. of7'-hydroxymethylzearalane in 5 m1. of dry pyridine. Cool the mixture onan ice bath and add with stirring 0.76 g. of -toluenesulfonylchloride.Allow the mixture to stand at room temperature for 15 hrs. and pour into50 m1. of ice water. Separate the insolubles by filtration and dissolvein chloroform. Wash the chloroform solution with dilute hydrochloricacid and saturated brine. Dry the washed solution over magnesium sulfateand remove the solvent in vacuo.

Take up the residue obtained above in 5 ml. of dimethylformamide and add0.32 g. of potassium cyanide. Heat the mixture on a steam bath for 24hrs., cool and pour into 50 ml. of water. Separate the insolubles byfiltration and dissolve in chloroform. Wash the chloroform solution fourtimes with water, dry over magnesium sulfate and remove the solvent invacuo.

Dissolve the residue obtained above in 10 ml. of absolute methanol andbubble hydrogen chloride through the mixture at temperatures below 40 C.with stirring for 30 mins. Reflux the mixture for 90 mins. and thendistill away 8 ml. of methanol. Add 10 ml. of water and stir at roomtemperature for 15 hrs. Extract the reaction mixture with chloroform,wash the chloroform extract with water, dry over magnesium sulfate andremove the solvent in vacuo. Purify the residue by thinlayerchromatography on four 1,000 silica gel plates using 10 percent ethylacetate in hexane as an eluant to obtain 7'-carbomethoxymethylzearalane.

EXAMPLE 7 7'-Carboxymethylzearalane Dissolve 0.38 g. of7-carbomethoxymethylzearalane in .5 ml. of 0.1N sodium hydroxide. Warmthe mixture on a steam bath for 1 hr., cool and acidify withhydrochloric acid. Collect the insolubles on a filter and purify bythin-layer chromatography on four 1,000u silica gel plates using 10percent methanol in chloroform as an eluant to obtain7-carboxymethylzearalane.

EXAMPLE 8 -Nitro-7"Carboxyzearalane To gm. of 7'-carboxyzea.ralane, add10 ml. of cool (10 C.) concentrated nitric acid. Stir the mixture at C.for 1 hr. and pour into 100 ml. of ice water. Collect the insolubles ona filter. Purify by recrystallization from nitromethane to obtain5-nitro-7-carboxyzearalane.

EXAMPLE 9 5-Nitro-7'-l-lydroxymethyzearalane Repeat the process ofexample 5 substituting an equivalent quantity of5-nitro-7'carboxyzearalane for the 7-forrnyl- Zearalane to obtain5-nitro-7'-hydroxymethylzearalane.

EXAMPLE l0 5-Nitro-7'-Formylzearalane Repeat the process for example 4substituting an equivalent quantity of 7'-hydroxymethylzearalane for the7-formylzearalane to obtain 5-nitro-7'-formylzearalane.

EXAMPLE 1 l 5-Nitro7'-Carboxymethylzearalane Repeat the process ofexample 8 substituting an equivalent quantity of7-carboxymethylzearalane for the 7'-formylzearalane to obtain5-nitro-7-carboxymethylzeralane.

EXAMPLE l2 2-Desoxy-7-Hydroxymethylzearalane A. 7410 -Formylzearalane4-Tetrahydropyranyl Ether Add 3 g. of 7'-formylzearalane to 8 ml. ofdihydropyran and 4 drops of concentrated hydrochloric acid. Stir 4 hrs.at room temperature and add an additional 8 ml. of dihydropyran. Stirfor an additional 48 hrs. at room temperature. Remove excessdihydropyran in vacuo and purify the residue by column chromatography on200 gm. of silica gel using 10 percent ethyl acetate in benzene as aneluant to obtain 7-formylzearalane 4-tetrahydropyranyl ether.

B. 7'-Formylzearalane 2-(l-Phenyltetrazolyl) Tetrahydropyranyl Ether Toa mixture of 1.33 g. of 7'-formylzearalane 4- tetrahydropyranyl ether,0.54 g. of l-phenyl-S-chlorotetrazole and 0.83 g. of anhydrous potassiumcarbonate, add 5 ml. of acetone. Reflux and stir the mixture undernitrogen for 3 days. Filter the mixture and remove the solvent from thefiltrate in vacuo. Purify by column chromatography on silica gel 100 g.)using 10 percent ethyl acetate in benzene as the Ether-4- eluant toobtain 7'-fonnylzearalane 2-( l-phenyl-tetrazolyl)ether-4-tetrahydropyranyl ether.

C. 2-Desoxy-7'-Hydroxymethylzearalane Shake l. l g. of the dietherobtained instep C above with 200 mg. of 5 percent platinum-on-carboncatalyst in 200 ml. of 95 percent ethanol for 48 hrs. at 70 C. under 50p.s.i. hydrogen pressure. Remove the insolubles by filtration andevaporate the solvent from the filtrate in vacuo. Add 50 ml. ofchloroform and b 1 ml. of concentrated hydrochloric acid to the residue.Stir and warm the mixture on a steam bath for 15 mins. Remove thesolvent in vacuo. Purify by column chromatography on 200 mg. of basicalumina (MERCK & CO.. Inc., Rahway, New Jersey, USA.) eluting first withethers to remove phenyltetrazole and then with l0 percent methanol inchloroform to obtain 2-desoxy-7-7-hydroxymethylzearalane.

EXAMPLE 1 3 2-Desoxy-7'-Fonnylzearalane Repeat the process of example 4substituting an equivalent quantity of2-desoxy-7'-hydroxymethylzearalane for the 7'- formylzearalane in orderto obtain 2-desoxy-7-formylzearalane.

EXAMPLE 14 2-Desoxy-7-Carboxyzearalane Repeat the process of example 4substituting an equivalent quantity of2-desoxy-7'-hydroxymethylzearalane for the 7'- formylzearalane and using0.5 ml. of Jones Reagent in order to obtain 2-desoxy-7'-carboxyzearalane.

EXAMPLE [5 7'-Formylzearalane 2,4-Dimethyl Ether Add 5 ml. of dimethylsulfate to a solution of 2 g. of 7-formylzearalane in ml. of 10 percentsodium hydroxide and 20 ml. of water. Stir the mixture for 30 min. at 20C. and add an additional 5 ml. of dimethyl sulfate. Stir for anadditional min. at b 25 C. Separate the solid precipitate by filtration,wash with water and dry. Purify by recrystallization from ethanol toobtain 7'-formylzearalane 2,4-dimethyl ether.

By substituting an equivalent quantity of a diaklyl or a diaralkylsulfate such as, for example, diethyl sulfate, diisopropyl sulfate,diamyl sulfate, dihexylsulfate and dibenzyl sulfate for the dimethylsulfate employed in the process described above, the corresponding2,4-diethyl, 2,4- diisopropyl, 2,4-diamyl, 2,4-dihexyl and 2,4-dibenzylether derivatives of 7-formylzearalane can be prepared.

EXAMPLE l6 7'-Formylzearalane Z-Methyl Ether Acidify the filtrateobtained after the separation of the precipitate from the reactionmixture of example 15 with 25 ml. of 12 N sulfuric acid. Separate theprecipitate by filtration, wash with water and dry. Purify therecrystallization from a mixture of ethanol and water to obtain7'-formylzearalane 2- methyl ether.

EXAMPLE l7 7'-Formylzearalane 4-Benzyl Ether To a stirred solution of480 mg. of 7'-formyl 2,4-dibenzyl ether in 5 ml. of methylene chlorideunder a nitrogen atmosphere at 0 C. add a cold solution of 2 ml. ofboron tribromide in 2 ml. of methylene chloride. Pour the reactionmixture immediately onto 50 ml. of crushed ice with stirring. Make thereaction mixture basic with potassium bicarbonate and extract withmethylene chloride. Wash the extract with saturated brine, dry overmagnesium sulfate and concentrate to dryness in vacuo. Purify byrecrystallization from nitromethane to obtain 7'-formylzearalane4-benzyl ether.

By substituting 7-formylzearalane 2,4-dialkyl (or diaralkyl) ethers suchas for example, the 2,4-dimethyl 2,4-diethyl, 2,4- dibutyl and2,4-dihexyl ethers for the dibenzyl ether employed EXAMPLE l87'-Formylzearalane 2,4-Diacetate Dissolve 24 g. of 7'-formylzearalane in150 ml. of pyridine and add l4.4 ml. of acetic anhydride. Stir for 12hrs. at room temperature. Pour the reaction mixture into 1,000 ml. ofice water and extract with 500 ml. of ether. Wash the ether extract withdilute hydrochloric acid and with water. Dry the washed extract overmagnesium sulfate and concentrate to dryness. Recrystallize from ethanolto obtain 7'-formylzearalane 2,4-diacetate.

By substituting an alkanoyl or aralkenoyl anhydride such as, forexample. propionic anhydride. butyric anhydride. hexanoic anhydride andbenzoic anhydride for the acetic anhydride employed in the above processthe corresponding 2.4- dipropionate, 2,4-dibutyrate. 2,4-dihexanoate and2,4- dibenmate ester derivatives of 7'-formylzearalane can be prepared.

EXAMPLE l9 7'-Formylzearalane Z-Methyl Ether-4-Acetate Dissolve 12 g. of7-formylzearalane 2-methyl ether in 80 ml. of pyridine and add 8 ml. ofacetic anhydride. Stir for 12 hrs. at room temperature Pour the reactionmixture into 500 ml. of ice water and extract with 500 ml. of ether.Wash the ether extract wit dilute hydrochloric acid and with water. Drythe ether extract over magnesium sulfate and concentrate to dryness.Recrystallize from ethanol to obtain 7'-formylzearalane Z-methylether-4-acetate.

EXAMPLE 20 7'-Formylzearalane 4-Ethyl Ether -2-Benzoate Repeat theprocess for example 19 substituting an equivalent quantity of7'-formylzearalane 4-ethyl ether for the 7'-formylzearalane Z-methylether and an equivalent quantity of benzoic anhydride for the aceticanhydride to obtain 7'-formylzearalane 4-ethyl ether-Z-benzoate.

EXAMPLE 2] EXAMPLE 22 7'-Formylzearalane Z-Benzoate Repeat the processof example 2l substituting an equivalent quantity of 7'-formylzearalane4-ethyl ether-2-benzoate for the 7'formylzearalane Z-methyl etherd-acetate to obtain 7'- formylzearalane 2-benzoate.

EXAMPLE 23 7'-Formylzearalane Z-Methyl Ether-4-Ethyl Ether To a solutionof 21;. of 7'-formylzearalane 2-methyl ether 80 ml. of percent potassiumhydroxide and ml. of water, add 5 ml. of diethyl sulfate. Stir themixture for mins. at 20 C. and allow the mixture to warm to 25 C. withcontinued stirring for an addiitonal 90 mins. Separate the precipitateby filtration, wash with water and dry. Recrystallize from ethanol toobtain 7'-formylzearalane 2-methyl ether-4-ethyl ether.

1Q. EXAMPLE 24 7'-Formylzearalane b 2-Acetate-4-Propionate Dissolve 6 g.of 7'-formylzearalane 2-acetate in 40 ml. of pyridine and add 4 ml. ofpropionic anhydride. Stir for 12 hrs. at room temperature. Pour thereaction mixture into 250 ml. of ice water and extract with 300 ml. ofether. Wash the ether extract with dilute hydrochloric acid and withwater. Dry the washed extract over magnesium sulfate and concentrate todryness. Recrystallize from methanol to obtain 7'-formylzearalane2-acetate-4-propionate.

EXAMPLE 25 2Desoxy-7'-Formylzearalane 4-Methyl Ether Repeat the processfor example 15 substituting an equivalent quantity of2-desoxy-7'-formylzearalane for the 7'- formylzearalane to obtain2desoxy-7'-formylzearalane 4- methyl ether.

EXAMPLE 26 2-Desoxy-7'-Carboxyzearalane 4-Methyl Ether Repeat theprocess of example 4 substituting an equivalent quantity of2-desoxy-7'-formylzearalane 4-methyl ether for the 7'-formylzearalane toobtain 2-desoxy-7-carboxy zearalane 4-methyl ether.

EXAMPLE 27 2-Desoxy-7-Hydroxymethylzearalane 4-Methyl Ether Repeat theprocess of example 5 substituting an equivalent quantity of2-desoxy-7'-fonnylzearalane to obtain 2-desoxy-7'-hydroxymethylzearalane 4-methyl ether.

Although applicants have described their invention above wit referenceto particular embodiments, many modifications will suggest themselves tothose skilled in the art from a study of the foregoing description.Applicants consider all such modifications to be the full equivalent ofthe embodiments specifically described and to fall within the scope oftheir invention.

The subject matter which applicants regard as their invention isparticularly pointed out and distinctly claimed as follows.

What is claimed is:

1. A compound of the formula:

wherein A is a member selected from the group consisting of hydrogen,hydroxy and -OR; R is a member of the group consisting of hydrogen,loweralkyl, benzyl, loweralkanoyl and benzoyl; B is a member of thegroup consisting of hydrogen and nitro, provided that B cannot be nitrowhen A is hydrogen; and R is a member of the group consisting of -CH OH,CHO, -COOR" and CH -COOR" wherein R" is a member of the group consistingof hydrogen, loweralkyl and benzyl.

2. A compound of claim 1 wherein A is hydrogen.

3. A compound of claim 1 wherein B is nitro.

4%. A compound of claim 2 wherein R is CH OH. 115.7'-Carboxymethylzearalane. 5. A compound of claim 2 wherein R is -CH().116. A process for preparing 7'-formylzearalane which com- 6. A compoundof claim 2 wherein R is 'COOR" and R" is prises: a member selected fromthe group consisting of hydrogen, a. treating zearalenone 2,4-dibenzylether with ethyl forloweralkyl and benzyl. mate in the presence of astrong base in order to obtain A mp und of Claim 2 wherein R is CH CO0R"7-hydroxymethylenezearalenone 2,4-dibenzyl ether; and

and R" is a member selected from the group consisting of t ting Said7'-hydroxymethylenezearal non hydrogen, lowemlkyl, and benzyL dibenzylether with cyclohexanol to form the correspond- 8. A compound of claim 3wherein R is CH OH. t; 7563181; and 9 A compound f l i 3 h i R i CH0 [0c. treating said 7-ether intermediate with a reducing agent 10. A om ndf l i 3 h i R i 400 and followed by treatment of the residue with astrong mineral is a member selected from the group consisting ofhydrogen, acid obtain T y y loweralkyl and benzyl. dibenzylether; d I

ll. A compound of claim 3 wherein R is -CH2-C0OR" y r s s salt} 6 y y eand R is a member Selected from the group consisting of 2,4-tlilbenzylether m the presence of a hydrogenation cata yst.

hydrogen, loweralkyl and benzyl.

l2. 7'-Formylzearalane. l3. 7'-Hydroxymethylzearalane. l4.7'-Carboxyzearalane.

117. The process of claim 17 wherein the reducing agent is sodiumborohydride.

I? I I HI

2. A compound of claim 1 wherein A is hydrogen.
 3. A compound of claim 1wherein B is nitro.
 4. A compound of claim 2 wherein R'' is -CH2OH.
 5. Acompound of claim 2 wherein R'' is -CHO.
 6. A compound of claim 2wherein R'' is -COOR'''' and R'''' is a member selected from the groupconsisting of hydrogen, loweralkyl and benzyl.
 7. A compound of claim 2wherein R'' is -CH2-COOR'''' and R'''' is a member selected from thegroup consisting of hydrogen, loweralkyl, and benzyl.
 8. A compound ofclaim 3 wherein R'' is -CH2OH.
 9. A compound of claim 3 wherein R'' is-CHO.
 10. A compound of claim 3 wherein R'' is -COOR'''' and R'''' is amember selected from the group consisting of hydrogen, loweralkyl andbenzyl.
 11. A compound of claim 3 wherein R'' is -CH2-COOR'''' and R''''is a member selected from the group consisting of hydrogen, loweralkyland benzyl.
 12. 7''-Formylzearalane.
 13. 7''-Hydroxymethylzearalane. 14.7''-Carboxyzearalane.
 15. 7''-Carboxymethylzearalane.
 16. A process forpreparing 7''-formylzearalane which comprises: a. treating zearalenone2,4-dibenzyl ether with ethyl formate in the presence of a strong basein order to obtain 7''-hydroxymethylenezearalenone 2,4-dibenzyl ether;and b. treating said 7''-hydroxymethylenezearalenone 2,4-dibenzyl etherwith cyclohexanol to form the corresponding 7''-ether; and c. treatingsaid 7''-ether intermediate with a reducing agent followed by treatmentof the residue with a strong mineral acid to obtain6''(7'')-dehydro-7''-formylzearalene 2,4-dibenzyl ether; and d.hydrogenating said 6''(7'')-dehydro-7''-formylzearalene 2,4-diBenzylether in the presence of a hydrogenation catalyst.
 17. The process ofclaim 17 wherein the reducing agent is sodium borohydride.